Recombinant human MBD4

Catalog # : EPX-002-RBV

Source : Human

Expressed in : SF9 cells

Quantity : 10µg of enzymatically active recombinant MBD4 at 0.5µg /µl

Background:

MBD4, methyl-CpG-binding domain protein 4, is a mismatch-specific DNA N-glycosylase involved in DNA repair (1). Has thymine glycosylase activity and is specific for G:T mismatches within methylated and unmethylated CpG sites (2). Can also remove uracil or 5-fluorouracil in G:U mismatches. Interacts with MLH1.

Protein details:

Recombinant human N-terminal His tagged MBD4 was produced in SF9 insect cells, purified using FPLC and formulated in a storage buffer containing 20mM Tris-Cl pH 7.6, 1mM EDTA, 0.15 M NaCl, 10% glycerol, 0.5mM PMSF and 1mM DTT. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

Quality control:

Each lot has been evaluated by 10% Tris Glycine SDS-PAGE.

SDS-PAGE gel of recombinant His tagged MBD4 (Lane 2).  Lane 1, protein molecular weight marker.		G/T mismatch containing DNA incubated (lane 2) or not (lane 1) with recombinant MBD4 and run on PAGE under denaturing conditions. Note that a 35 bp cut product was generated in the presence of MBD4.

In vitro glycosylase assay:

A 70 bp DNA containing a G/T mismatch was incubated or not with recombinant MBD4 for 30 minutes at 37°C. The reaction products were treated with NaOH and run on PAGE under denaturing conditions. Note that a 35 bp cut product was generated in the presence of MBD4.

Storage:

-80°C

Guarantee:

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

Purity:

>98% purity by SDS PAGE.

Sequence:

MBD4 (Mw: 66,987 Da.)
MIHHHHHHGTTGLESLSLGDRGAAPTVTSSERL
VPDPPNDLRKEDVAMELERVGEDEEQMMIKRSS
ECNPLLQEPIASAQFGATAGTECRKSVPCGWER
VVKQRLFGKTAGRFDVYFISPQGLKFRSKSSLA
NYLHKNGETSLKPEDFDFTVLSKRGIKSRYKDC
SMAALTSHLQNQSNNSNWNLRTRSKCKKDVFMP
PSSSSELQESRGLSNFTSTHLLLKEDEGVDDVN
FRKVRKPKGKVTILKGIPIKKTKKGCRKSCSGF
VQSDSKRESVCNKADAESEPVAQKSQLDRTVCI
SDAGACGETLSVTSEENSLVKKKERSLSSGSNF
CSEQKTSGIINKFCSAKDSEHNEKYEDTFLESE
EIGTKVEVVERKEHLHTDILKRGSEMDNNCSPT
RKDFTGEKIFQEDTIPRTQIERRKTSLYFSSKY
NKEALSPPRRKAFKKWTPPRSPFNLVQETLFHD
PWKLLIATIFLNRTSGKMAIPVLWKFLEKYPSA
EVARTADWRDVSELLKPLGLYDLRAKTIVKFSD
EYLTKQWKYPIELHGIGKYGNDSYRIFCVNEWK
QVHPEDHKLNKYHDWLWENHEKLSLS

References:

1.    Bellacosa et al., Proc. Natl. Acad. Sci. U.S.A. 96:3969-3974(1999)
2.    Petronzelli et al., J. Biol. Chem. 275:32422-32429(2000)